Determination of the unbound free fatty acid profiles in biological and industrial specimens
AssigneeAlan Kleinfeld
InventorsAlan Kleinfeld, Andrew Huber
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Abstract
The invention relates to determination of the profile of unbound free fatty acids (FFAu) in biological samples, such as human and animal blood specimens and plant and animal oils, by measuring the fluorescence response of sets of different fluorescently labeled fatty acid binding proteins (probes) that undergo a change in fluorescence ratio at 2 wavelengths upon binding an FFAu. Use of these profiles in human and animal disease, in basic research, in drug development and in industrial uses is disclosed.
Claims19
A method for identifying and measuring an amount of one or more unbound free fatty acids (FFAu) in a biological sample comprising the steps of:
- (a) providing a set of probes comprising a plurality of FFAu probes wherein each probe in the plurality of FFAu probes has a specificity for a plurality of FFAu of interest that is different from and complementary to the specificity of one or more other probes in the set of probes, and wherein at least 60% of probes in the set of probes is labeled at a cysteine residue;
- (b) combining each individual probe from the plurality of FFAu probes with a separate aliquot of the biological sample for each individual probe;
- (c) measuring a fluorescence index at two different emission wavelengths for each probe in the combined probe and aliquot of the biological sample from step (b);
- (d) calculating the ratio of the fluorescence index at the two wavelengths from step (c) for each probe;
- and
- (e) determining an identity and measuring an amount of the one or more FFAu present in the biological sample based on the calculated ratio of fluorescence index for each probe,
- wherein the set of probes comprises more probes than the number of FFAu to be identified and measured.
Dependencies are inferred from the claim text.